国际生殖健康/计划生育杂志

国际生殖健康/计划生育 ›› 2015, Vol. 34 ›› Issue (5): 357-360.

• 论著 •    下一篇

冷冻保存对人类卵母细胞线粒体DNA缺失和能量代谢的影响

蔡学泳#，彭南妮，刁瑞英，李凌伟，陈燕   

  1. 518035 深圳市第二人民医院生殖医学科（蔡学泳，刁瑞英，陈燕），深圳罗湖区人民医院生殖中心（彭南妮，李凌伟）
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2015-09-15 发布日期:2015-09-15
  • 通讯作者: 蔡学泳

Study on Human Oocyte Mitochondrial DNA Deletions and Energy Metabolism before and after Cryopreservation

CAI Xue-yong, PENG Nan-ni, DIAO Rui-ying, LI Ling-wei, CHEN Yan   

  1. Reproductive Centre, Shenzhen Secondary People Hospital, Shenzhen 518035，China（CAI Xue-yong, DIAO Rui-ying, CHEN Yan）；Reproductive Centre, Shenzhen Luo Hu People Hospital, Shenzhen 518030, China（PENG Nan-ni, LI Ling-wei）
  • Received:1900-01-01 Revised:1900-01-01 Published:2015-09-15 Online:2015-09-15
  • Contact: CAI Xue-yong

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摘要： 目的：通过检测玻璃化冷冻前后卵母细胞线粒体DNA（mtDNA）缺失和能量代谢变化，以探讨冷冻保存对卵母细胞质量的影响。方法：收集体外受精（IVF）废弃的人卵母细胞和受精卵，包括未受精（0PN）、两原核（2PN）和三原核（3PN）；随机分为2组，A组为经玻璃化冷冻保存，B组为新鲜对照组。采用巢式聚合酶链反应（nest PCR）检测冷冻前后卵母细胞mtDNA缺失差异；并进一步比较冷冻前后卵母细胞mtDNA拷贝数和三磷酸腺苷（ATP）含量变化。结果：卵母细胞mtDNA缺失率和mtDNA拷贝数在冷冻处理前后差异无统计学意义（P＞0.05）；冷冻后卵母细胞内ATP含量低于冷冻前新鲜卵母细胞。结论：冷冻保存对卵母细胞mtDNA缺失及拷贝数影响较小，可较好地实现保存卵母细胞的目的。

关键词: 低温保存, 卵母细胞, DNA, 线粒体, 应激, 氧化性

Abstract: Objective: To explore the effect of cryopreservation on the mitochondrial DNA（ｍｔＤＮＡ） deletions and energy metabolism of human oocyte, so as to investigate the effect of cryopreservation on oocytes preservation. Methods: The dumped oocytes with 0PN, 2PN and 3PN during routine IVF were collected for this cryopreservation study. Those oocytes were randomly divided into two groups, one for vitrification cryopreservation （Ｇｒｏｕｐ Ａ） and the other as control （Group B, without cryopreservation）. The nested polymerase chain reaction （ＰＣＲ） was used to detect the mtDNA deletions of oocyte mtDNA before and after cryopreservation. Real-time quantitative PCR was adopted to determine the copy number of oocyte mtDNA. ATP contents were detected using luciferin-luciferase assays. Results: There were not significant differences in the mutation frequency of oocyte mtDNA deletions and the copy number of mtDNA between two groups（P＞0.05）. However, The ATP content in the group A was significantly lower than that in the group B. Conclusions: The effect of cryopreservation on the oocyte mtDNA deletions and the copy number of mtDNA was limited, suggesting that cryopreservation can be properly used to preserve oocyte.

Key words: Cryopreservation, Oocytes, DNA, mitochondrial, Oxidative stress